Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

New lambda and plasmid vectors for expression cloning in mammalian cells

G Del Sal1, G Manfioletti, S Gustincich

  • 1Laboratorio Nazionale Consorzio Interuniversitario per le Biotecnologie, Trieste, Italy.

Biotechniques
|January 1, 1994
PubMed
Summary

Researchers developed new lambda phage and plasmid cloning vectors for efficient gene cloning and expression. These vectors facilitate cDNA transfer and functional screening in mammalian cells, improving research workflows.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The growth suppressing gas1 product is a GPI-linked protein.

FEBS letters·2000
Same author

Analysis of the domain requirement in Gas1 growth suppressing activity.

FEBS letters·2000
Same author

Proof of genetic heterogeneity in the proximal myotonic myopathy syndrome (PROMM) and its relationship to myotonic dystrophy type 2 (DM2).

Neuromuscular disorders : NMD·2000
Same author

Hyperparathyroidism in a patient with proximal myotonic myopathy (PROMM).

Neuromuscular disorders : NMD·2000
Same author

Large vasodilatations in skeletal muscle of resting conscious dogs and their contribution to blood pressure variability.

The Journal of physiology·2000
Same author

Cell-cycle regulation of the p53-inducible gene B99.

FEBS letters·2000

Area of Science:

  • Molecular Biology
  • Gene Cloning and Expression

Background:

  • Efficient cloning vectors are crucial for molecular biology research.
  • Existing vectors may have limitations in cDNA insertion capacity and screening methods.

Purpose of the Study:

  • To construct and characterize a novel family of lambda phage and plasmid cloning vectors.
  • To enable efficient cDNA cloning, sequencing, in vitro transcription/translation, and functional expression screening.

Main Methods:

  • Construction of lambda GDST3/T7 phage vector with an expanded multiple cloning site (MCS).
  • Insertion of MCS into eukaryotic expression plasmids pGDSV3 and pGDSV7 in both orientations.
  • Utilized T3 and T7 promoters for direct sequencing and in vitro applications.
  • Employed SfiI restriction sites for seamless cDNA transfer between vectors.

Related Experiment Videos

Main Results:

  • Lambda GDST3/T7 accommodates cDNA inserts up to 14 kb.
  • The MCS provides eight unique restriction sites, including asymmetrical SfiI sites.
  • Vectors facilitate high signal-to-noise ratio screening using nucleic acid or antibody probes.
  • Efficient transfer of cDNAs into expression plasmids for mammalian cell screening was demonstrated.

Conclusions:

  • The new lambda phage and plasmid vectors offer enhanced capabilities for molecular cloning and functional studies.
  • These vectors streamline the process from primary screening to functional secondary screening in mammalian cells.
  • The described system provides a versatile tool for gene discovery and characterization.